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149]

Original Article

Effect of 17% ethylenediaminetetraacetic acid

and 0.2% chitosan on pushout bond strength of
biodentine and ProRoot mineral trioxide aggregate:
An in vitro study
Pandi Prasanthi, Roopadevi Garlapati, Bolla Nagesh, Varri Sujana, K. Madhu Kiran Naik, Bandaru Yamini
Department of Conservative Dentistry and Endodontics, Sibar Institute of Dental Sciences, Guntur, Andhra Pradesh, India

Abstract
Aim: The purpose of this study was to evaluate the effect of 17% ethylenediaminetetraacetic acid (EDTA) and 0.2% chitosan on
pushout bond strength of biodentine and ProRoot mineral trioxide aggregate (MTA).
Materials and Methods: Midroot dentin of single‑rooted human canine teeth were sectioned into 2‑mm‑thick slices
horizontally (n = 60). The canal space of each dentin slice was enlarged with a 1.3‑mm‑diameter diamond bur. The samples
were divided into two groups (n = 30) based on the type of perforation repair material placed, i.e., Biodentine and ProRoot
MTA. The samples were wrapped in wet gauge for 10 min, and based on the type of chelating agent used for removal of smear
layer, each group is further divided into three subgroups (n = 10), to be immersed into saline (control), 17% EDTA and 0.2%
chitosan for 30 min, and a wet cotton pellet was placed over each test material. After 48 h of incubation, the dislodgement
resistance of the samples was measured using a universal testing machine.
Statistical Analysis: Data were analyzed using one‑way analysis of variance and post hoc Tukey tests. The level of statistical
significance was set at 0.05.
Results: Biodentine showed significantly higher pushout bond strength than ProRoot MTA. Biodentine and ProRoot MTA lost
strength when exposed to 0.2% chitosan.
Conclusion: Biodentine showed considerable performance as a perforation repair material than ProRoot MTA even after being
exposed to various endodontic chelating agents.
Keywords: Biodentine; chelating agents; mineral trioxide aggregate; pushout bond strength

INTRODUCTION which usually prognosticates a high failure risk. Hence,

immediate repair of perforation is important to avoid
Perforation is a procedural complication that can occur contamination and thereby preventing endodontic
during endodontic therapy or post space preparation failure.[2]
of teeth.[1] Delayed perforation repair therapy can cause
periodontal or endodontic lesions and lateral periodontal An ideal endodontic root repair material should be
abscesses occurring secondary to delayed diagnosis, biocompatible, radiopaque, antibacterial, dimensionally
stable, easy to manipulate, and unaffected by blood
Address for correspondence: contamination.[3] It should also remain in place under
Dr. Pandi Prasanthi, Department of Conservative Dentistry and dislodging forces, such as mechanical loads of occlusion
Endodontics, Sibar Institute of Dental Sciences, Takkellapadu,
Guntur 522 509, Andhra Pradesh, India.
E‑mail: dr.rupagarlapati@gmail.com This is an open access journal, and articles are distributed under the terms
Date of submission : 02.02.2019 of the Creative Commons Attribution‑NonCommercial‑ShareAlike 4.0
Review completed : 18.03.2019 License, which allows others to remix, tweak, and build upon the work
Date of acceptance : 31.07.2019 non‑commercially, as long as appropriate credit is given and the new
creations are licensed under the identical terms.
Access this article online
For reprints contact: reprints@medknow.com
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Website:
www.jcd.org.in How to cite this article: Prasanthi P, Garlapati R, Nagesh B,
Sujana V, Kiran Naik KM, Yamini B. Effect of 17%
ethylenediaminetetraacetic acid and 0.2% chitosan on pushout bond
DOI:
strength of biodentine and ProRoot mineral trioxide aggregate: An
10.4103/JCD.JCD_56_19
in vitro study. J Conserv Dent 2019;22:387-90.

© 2019 Journal of Conservative Dentistry | Published by Wolters Kluwer - Medknow 387

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Prasanthi, et al.: Effect of 17% EDTA and 0.2% chitosan
or the condensation of restorative materials over it.
Therefore, the pushout bond strength is an important
factor for perforation repair materials.[4]
Although many perforation repair materials such as
amalgam, cavity, composite resin, glass ionomer cement,
calcium hydroxide, super EBA, intermediate restorative
material, and mineral trioxide aggregate (MTA) have
been used, most of these materials show significant
shortcomings in one or more of the following areas:
solubility, leakage, biocompatibility, handling properties,
and moisture incompatibility.[1] Despite the numerous
favorable properties of MTA that support its clinical use
when compared with other traditional materials, there are
several critical drawbacks of MTA such as the prolonged
setting time, difficult handling characteristics, high cost,
and potential of discoloration. A variety of new calcium
silicate‑based materials have been developed recently
aiming to improve MTA shortcomings.[1]
Biodentine is introduced as an alternative to MTA.
Biodentine (septodont) is a high‑purity calcium silicate‑based
dental material composed of tricalcium silicate, calcium
carbonate, zirconium oxide, and a water‑based liquid
containing calcium chloride as the setting accelerator and
water‑reducing agent.[5]
After repairing the furcal perforation, endodontic
treatment should be performed with various irrigants
to disinfect the root canal system, including chelating
agents for smear layer removal.[6] During biomechanical
preparation, the smear layer is formed over the cut
dentinal surfaces.[7] Various agents, such as sodium
hypochlorite, ethylenediaminetetraacetic acid (17% EDTA),
mixture of tetracycline acid detergent, and organic acids
(e.g., citric acid), have been introduced for smear layer
removal.[7] The alternating use of 17% EDTA and sodium
hypochlorite has been recommended for the efficient
removal of the smear layer. However, the use of these
solutions may cause periapical inflammatory reactions and
reduce periapical healing.[7] 17% EDTA, which is the most
commonly used calcium chelator, has inhibited the setting
of MTA.[8] To minimize their harmful effects on periapical
tissues, the use of biocompatible solutions is essential.[7]
Chitosan is a natural polysaccharide that is biocompatible,
nontoxic, and having chelating property.[7] Chitosan is
natural polysaccharide obtained by the deacetylation of
chitin, but it has limited solubility. Chitosan and chitin do
not cause any biological hazard, and both are inexpensive.
Chitosan exhibits many biological actions such as
antimicrobial, wound healing, mucoadhesive, sustained
drug releasing property. Chitosan is used as as a chelating
agent and as an irrigating solution during endodontic
treatment.[7]
388 Journal of Conservative Dentistry | Volume 22 | Issue 4 | July-August 2019
Hence, in this study, smear layer removing agents 17%
EDTA and 0.2% chitosan were used. Root canal treatment
procedure may cause unavoidable contact of chelating
solutions with the repair materials. Thus, the purpose of
this in vitro study was to evaluate the effect of chelating
agents, i.e., 17% EDTA, 0.2% chitosan on the pushout bond
strength of biodentine in comparison with ProRoot MTA.
MATERIALS AND METHODS
Freshly extracted single‑rooted human canine teeth were
selected. The crowns of all teeth were removed, and the
midroot dentin was sectioned horizontally into slices with
a thickness of 2 mm (n = 60) using hard tissue microtome.
In each slice, the space of canal was enlarged with a
1.3‑mm‑diameter diamond bur. The specimens (n = 60)
were randomly divided into two groups (n = 30) and the
following test materials were used: Group 1: Biodentine
and Group 2: ProRoot MTA. The test materials were
incrementally placed into canal spaces of the dentin slices
and condensed. Excess material was trimmed from the
surface of the samples with a scalpel. Subsequently, the
samples were wrapped in wet gauze placed in incubator
and allowed to set for 10 min at 37ºC with 100% humidity.
Setting time of biodentine is 10 min, so it was allowed
to set for 10 min. As the initial setting time of ProRoot
MTA is 10 min, it was allowed to set for 10 min only.
Immediately after incubation, the samples were divided
into three subgroups (n = 10) to be immersed into saline
solution (control), 17% EDTA, 0.2% chitosan. After 30 min
of immersion, all samples were removed from the test
solutions, rinsed with distilled water, and were placed in
incubator at 37ºC with 100% humidity for 48 h.
Pushout test
After 48 h, the pushout bond strength values
were measured using a universal testing machine
(Instron Universal Machine) [Figure 1]. The samples were
placed on a base of a metal slab of universal testing machine
to allow the free motion of the plunger. The compressive
load was applied by exerting a download pressure on the
surface of test material in each sample, with the Instron
probe moving at a constant speed of 1 mm/min. The plunger
size of 1 mm had a clearance of approximately 0.2 mm from
the margin of the dentinal wall to ensure contact only with
the test materials. The maximum force applied to materials
at the time of dislodgement was recorded in newtons. The
pushout bond strength in megapascal (MPa) was calculated
by dividing this force (N) by the surface area of the test
material where N/2p × r × h, p is the constant 3.14, r is the
root canal radius, and h is the thickness of the dentin slice
in millimeters. Data were analyzed using one‑way analysis
of variance and post hoc Tukey tests. The level of statistical
significant was set at P < 0.05.
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Prasanthi, et al.: Effect of 17% EDTA and 0.2% chitosan

RESULTS and medicaments into the dentinal tubules. There is a

Table 1 shows the mean values and standard deviations of
the pushout bond strength (MPa) of all groups. The lowest
pushout bond strength was observed in the ProRoot MTA
group (P < 0.05). Biodentine displayed a significantly
higher resistance to displacement than the ProRoot MTA
group. In biodentine, saline (control) group showed the
highest pushout bond strength whereas chitosan group
showed the least pushout bond strength next to EDTA
group. In ProRoot MTA, saline (control) group showed
the highest pushout bond strength, whereas chitosan
showed the least pushout bond strength next to EDTA
group [Figure 2].
DISCUSSION
After repair of the perforation, the success of the
endodontic therapy depends on a well‑placed coronal
restoration as well as the resistance of the repair material
to displacement forces happening while undergoing
condensation of permanent restorative materials. Thus,
the bond strength of the perforation repair materials is an
important factor in clinical practice.[1] To assess the bond
strength, the pushout bond test has been shown to be
efficient, practical, and reliable.[1,4]
The presence of smear layer may inhibit or significantly
delay the penetration of irrigating solutions, sealers,
controversy regarding the presence and removal of smear
layer. It is now generally advocated that the smear layer
should be removed prior to the root canal obturation to
facilitate better adaptation of the filling material to the
root canal wall and to improve adhesion.[5]
Alternate use of EDTA and NaOCl as retro smear layer
removing agents may cause periapical inflammatory
reactions at surgical site. Calt et  al. observed that usage
of EDTA for prolonged periods caused excessive tubular
and intertubular dentin erosion.[9] Hence, the use of
biocompatible retro smear layer removing agents is
essential. Chitosan is a natural polysaccharide that is
biocompatible, nontoxic, and having chelating property.[7]
This study evaluated the pushout bond strength between
biodentine and ProRoot MTA after exposure to endodontic
chelating agents, i.e., 17% EDTA and 0.2% chitosan.
In the present study, among ProRoot MTA groups,
saline‑treated ProRoot MTA samples resisted dislodgment
forces >17% EDTA‑treated ProRoot MTA samples and
0.2% chitosan‑treated ProRoot MTA samples. This was in
accordance with the results of the study done by Loxely
et al.  who reported that the compressive strength of MTA
increased when immersed in saline solution because of
the remaining unreacted mineral oxides. These may be
solidified after additional supplied hydration and may
result in the increased strength of material.[10]

Table 1: Mean pushout bond strength values and ProRoot MTA samples treated with 17% EDTA showed
standard deviations of all test groups lower resistance to dislodgement forces. Lee et al. showed
Groups Mean SD SE CV the adverse effects of 17% EDTA on calcium silicate‑based
Biodentin-Control 43.46 6.90 2.18 15.87
cement (CSC) hydration.[11] Yan et al. showed that 17% EDTA
Biodentin-Chitosan 12.41 4.19 1.32 33.77 decreased the bond strength between dentine and CSC.[12]
Biodentin-EDTA 16.66 4.37 1.38 26.23 Nagesh et  al. reported that sealing ability of MTA with
MTA-Control 20.37 3.31 1.05 16.27 chitosan was less.[7]
MTA-Chitosan 11.20 3.35 1.06 29.89
MTA-EDTA 15.94 1.98 0.63 12.44

Figure 2: Comparison of six groups (biodentine – control,

biodentine – chitosan, biodentine – ethylenediaminetetraacetic
acid, mineral trioxide aggregate – control, mineral trioxide
aggregate – chitosan and mineral trioxide aggregate –
Figure 1: Pushout bond test ethylenediaminetetraacetic acid)

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Prasanthi, et al.: Effect of 17% EDTA and 0.2% chitosan
Biodentine has a similar composition to MTA, differing
mostly by being aluminum‑free and having tantalum oxide as
a radiopacifier in place of the bismuth oxide. This is claimed
to be associated with improved biological property.[2] The
presence of calcium chloride, and the concordant reduced
setting time and contact time, probably provided the
high‑bond strength in biodentine group.[1]
Biodentine was more resistant to dislodgement forces
than MTA in the present study. The biomineralization
ability of biodentine, most likely through the formation of
tags, may be the reason of the dislodgement resistance.[1]
The higher bond strength values of biodentine may, in
part, result from its smaller particle size, which has the
potential to enhance penetration of the cement into the
medicament‑free dentinal tubules, leading to improved
bond strength.[13] This effect might be further reinforced
through the formation of dentinal bridges as a result
of crystal growth within the dentinal tubules, leading
to increased micromechanical retention (Han and Okiji
2011, Atmeh et  al.  2012).[14] Biodentine may have a
more prominent biomineralization ability than MTA,
as biodentine specimens showed wider Caand Sirich
dentine areas and larger incorporation depths than
MTA. This could be because of the amount of Ca and Si
dissolution that could be larger in biodentine than in
MTA. Chemical analysis of the interfacial dentine layer
confirmed increased Ca levels and Ca/P ratios in the
biodentine and MTA specimens.[15] This finding is related
to Ca incorporation. As such, the quality of the interfacial
dentine layer appeared to be improved. The higher
content of calcium‑releasing products in biodentine than
in MTA may promote to higher biomineralization and
higher bond strength.[16]
Biodentine also displayed a lower resistance to
dislodgement after exposure to 17% EDTA and chitosan
solutions. Lee et al. showed the adverse effects of 17% EDTA
on CSC hydration, microhardness, and cell adhesion. In
addition, Yan et al. showed that EDTA decreased the bond
strength between dentine and CSC. Uyanik et al. examined
the effect of different irrigation regimens on the sealing
ability of CSC and showed that using EDTA significantly
increases leakage of CSC.[17] In another study, CSC showed
significantly higher leakage when smear layer was removed
before placing the material.[3] This might be because of the
reaction between CSC and residual EDTA inside the root
canal.[3] Pushout bond strength is reduced when exposed to
chitosan due to more chelation property which interferes
with the setting reaction of CSCs.
CONCLUSION
Within the limitations of this in vitro study, it can be
concluded that:
390 Journal of Conservative Dentistry | Volume 22 | Issue 4 | July-August 2019
1. The force needed for dislodgement of biodentine from
root dentin was significantly higher than ProRoot MTA
2. Endodontic chelating agents influence the resistance
to dislodgement of biodentine and ProRoot MTA.
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
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